Characterization Of Allergens Research Articles

Neglected but Clinically Relevant Allergens in Korea.

Allergy diagnostics and immunotherapeutics in Asia heavily rely on imported products from Western countries, raising concerns about the accuracy and efficacy of these products for the management of Asian allergy patients. Recent advancements in allergen research have led to the identification and characterization of novel allergens from indigenous Korean species. While some allergens share homology with well-known allergens, others lack counterparts in imported allergen extracts. Classifying regional allergens in Asia into three categories based on their cross-reactivity with imported allergens offers valuable insights. Highly cross-reactive allergens, such as oak allergens Que m 1 from Quercus mongolica and Que ac 1 from Q. acutissima, can be effectively substituted with the imported allergens. Allergens with partial cross-reactivity, like the Asian needle ant allergen Pac c 3 (Antigen 5), permit limited diagnostic value by the currently available products. Unique allergens, including the Japanese hop allergen Hum j 6 (pectin methylesterase inhibitor) and the silkworm pupa allergen Bomb m 4 (30kDa hemolymph lipoprotein) lack alternatives in the available product list. Greater attention is needed, particularly for species listed as ecologically invasive in Western regions. Additionally, allergens from domestic fruits and vegetables causing pollen food allergy syndrome require characterization for the development of improved diagnostics.

Structural homology of mite profilins to plant profilins is not indicative of allergic cross-reactivity.

Structural and allergenic characterization of mite profilins has not been previously pursued to a similar extent as plant profilins. Here, we describe structures of profilins originating from Tyrophagus putrescentiae (registered allergen Tyr p 36.0101) and Dermatophagoides pteronyssinus (here termed Der p profilin), which are the first structures of profilins from Arachnida. Additionally, the thermal stabilities of mite and plant profilins are compared, suggesting that the high number of cysteine residues in mite profilins may play a role in their increased stability. We also examine the cross-reactivity of plant and mite profilins as well as investigate the relevance of these profilins in mite inhalant allergy. Despite their high structural similarity to other profilins, mite profilins have low sequence identity with plant and human profilins. Subsequently, these mite profilins most likely do not display cross-reactivity with plant profilins. At the same time the profilins have highly conserved poly(l-proline) and actin binding sites.

Characterization of the potential allergenicity of enzymatically hydrolyzed casein in Balb/c mouse model.

Bovine casein is a major allergen present in cow milk to induce anaphylaxis. In this study, the potential allergenicity of enzymatically hydrolyzed casein (HC) was evaluated based on in vitro and in vivo. The results showed that Alcalase and Protamex treatment (AT, PT) reduced the potential allergenicity of CN, with the greatest reductions of 68.25% and 50.75%, respectively. In addition, in vivo results showed that HC effectively alleviated allergic response symptoms of Balb/c mice; a significant tendency toward decreased serum IgG1 and mast cell tryptase levels was observed, accompanied by a decrease of Th2-associated IL-4, IL-5, and IL-13 and an increase of IFN-γ levels in spleen. Moreover, the inflammation of the lung, jejunum, and ileum was remarkably ameliorated. The findings indicated that HC induced a shift toward Th1 response and maintained the Th1/Th2 immune balance. Importantly, our results provide the basis for the production of hypoallergenic dairy products.

AKTUALNE PERSPEKTYWY ZASTOSOWANIA PROTEOMIKI DO IDENTYFIKACJI ORAZ CHARAKTERYSTYKI BIAŁEK POCHODZENIA OWADZIEGO WYKAZUJĄCYCH POTENCJAŁ ALERGIZUJĄCY

Currently, pet owners can easily access a broad array of commercial pet products specifically formulated to meet the unique nutritional needs of their cats and dogs. The trend towards organic pet foods, prioritizing whole ingredients over processed ones, is gaining more and more attention, emphasizing sustainability in sourcing dietary proteins. Insects, rich in essential amino acids, emerge as promising protein substitutes, though they do carry potential risks of triggering adverse food reactions, such as allergies in pets. This, in turn, provides a fertile ground for advanced proteomic analysis, facilitating comprehensive identification and quantification of allergens from edible insects. This includes defining immune epitopes originating from these allergens, thus fostering the discovery of potentially new proteins with allergenic potential and aiding in the in vitro diagnosis of allergen sensitization. This review presents the latest advancements in edible insect-derived allergens and the proteomic investigations conducted to analyse them. We offer insights into classical and advanced proteomic approaches designed for identifying and quantifying these allergens in various food matrices. Additionally, we focus on recent studies concerning the characterization of novel insect allergens, exploring sensitization and cross-reactivity mechanisms, with a specific emphasis on pets.

Identification and characterization of a novel nematode pan allergen (NPA) from Wuchereria bancrofti and their potential role in human filarial tropical pulmonary eosinophilia (TPE).

Tropical pulmonary eosinophilia (TPE) is a chronic respiratory syndrome associated with Lymphatic Filariasis (LF), a tropical parasitic infection of the human, transmitted by mosquitoes. The larval form of LF (microfilariae) are trapped in the lungs of TPE subjects have a major role in initiating the TPE syndrome. To date, there are no reports on the potential allergen that is responsible for generating parasite-specific IgE in TPE. In this project, we screened a cDNA expression library of the microfilarial stages of Wuchereria bancrofti with monoclonal IgE antibodies prepared from subjects with clinical filarial infections. Our studies identified a novel molecule that showed significant sequence similarity to an allergen. A blast analysis showed the presence of similar proteins in a number of nematodes parasites. Thus, we named this molecule as Nematode Pan Allergen (NPA). Subsequent functional analysis showed that NPA is a potent allergen that can cause release of histamine from mast cells, induce secretion of proinflammatory cytokines from alveolar macrophages and promote accumulation of eosinophils in the tissue, all of which occur in TPE lungs. Based on our results, we conclude that the NPA protein secreted by the microfilariae of W. bancrofti may play a significant role in the pathology of TPE syndrome in LF infected individuals. Further studies on this molecule can help design an approach to neutralize the NPA in an attempt to reduce the pathology associated with TPE in LF infected subjects.

Comprehensive characterization and detection of nut allergens in bakery foods using Q-TOF mass spectrometry and bioinformatics

Abstract Food allergy is a growing health issue worldwide and the demand for sensitive, robust and high-throughput analytical methods is rising. In recent years, mass spectrometry-based methods have been established for multiple food allergen detection. In the present study, a novel method was developed for the simultaneous detection of almond, cashew, peanut, and walnut allergens in bakery foods using liquid chromatography–mass spectrometry. Proteins unique to these four ingredients were extracted, followed by trypsin digestion, quadrupole time-of-flight (Q-TOF) mass spectrometry and bioinformatics analysis. The raw data were processed by de-novo sequencing module plus PEAKS DB (database search) module of the PEAKS software to screen peptides specific to each nut species. The thermal stability and uniqueness of these candidate peptides were further verified using triple quadrupole mass spectrometry (QQQ-MS) in multiple reaction monitoring (MRM) mode. Each nut species was represented by four peptides, all of which were validated for label-free quantification (LFQ). Calibration curves were constructed with good linearity and correlation coefficient (r2) greater than 0.99. The limits of detection (LODs) were determined to range from 0.11 to 0.31 mg/kg, and were compared with the reference doses proposed by Voluntary Incidental Trace Allergen Labelling (VITAL). The recoveries of the developed method in incurred bakery food matrices ranged from 72.5% to 92.1% with relative standard deviations (RSD) of <5.2%. The detection of undeclared allergens in commercial bakery food samples confirmed the presence of these allergens. In conclusion, this method provides insight into the qualitative and quantitative detection of trace levels of nut allergens in bakery foods.

Pork-cat syndrome

Meat allergy used to be considered a rare hypersensitivity reaction, but case reports of mammalian and poultry meat allergies became more common about 20 years ago. Allergies to pets such as cats (Felis domesticus) and dogs (Canis familiaris) are the second most common indoor allergic reactions in Western countries, affecting both adults and children. An important step in fur allergy diagnosis has been the introduction of molecular allergen-based diagnosis, which offers new opportunities for improved allergen characterization. An example of these new characterizations is the meat reactions of patients suffering from pork-cat syndrome. Patients with this syndrome sensitized to albumin from animal epithelia report symptoms after eating pork because there is cross-reactivity to albumins from different species. The management of a patient with an allergic condition to fur-bearing animals includes a detailed history of exposure to relevant allergens, the effect of exposure on symptoms and subsequent recommendations for controlled exposure, and in the case of pig-cat syndrome, the patient can tolerate well-cooked red meat, but not the insufficiently prepared one.

Yeast Surface Display Methodology for the Characterization of Food Allergens In Situ.

High throughput allergen characterization is often based on phage display technique which is limited by the constraints of a prokaryotic expression system such as potential loss of conformational epitopes and lack of post-translational modifications. Replacing the phage display platform with a yeast surface display system could accelerate the immunological characterization of complex structured allergens. Yeast surface display is a powerful technique that allows faster immunochemical characterization of allergens in situ without the need for protein purification. Yeast surface display offers an alternative that could lead to the improvement of standard immunodiagnostic and immunotherapeutic approaches. In this chapter, we describe a protocol on yeast surface display for the characterization of plant-derived food allergens using actinidin (Act d 1), a major kiwifruit allergen, as a model system.

Structural Characterization of Food Allergens by Nuclear Magnetic Resonance Spectroscopy.

As allergies, especially those triggered by food, are becoming more and more prevalent, it is of increasing importance to fully understand the structures and dynamic behaviors of allergenic proteins along with their interactions with potential natural ligands. Therefore, we have established a solid routine to achieve structural characterization of food allergens, especially for birch pollen-related cross-reactive proteins from the class 10 of pathogenesis-related proteins (PR-10), by nuclear magnetic resonance (NMR) spectroscopy. Following expression of the desired allergen in Escherichia coli in isotope-labeled minimal media, the three-dimensional solution structures of these proteins can be determined, and insight into ligand binding mechanics and structural dynamic properties are accessible through NMR spin relaxation experiments.

In Silico Identification and Characterization of Potential Red Seaweed Allergens

Red seaweeds such as Kappaphycus alvarezii and Porphyra yezoensis have many applications, especially in the food industry, which suggests the need for knowing their potential allergenicity. In silico approaches can be used to determine if a protein is an existing allergen or has the ability to cross-react with one. In this study, 318 sequences for Kappaphycus alvarezii and 641 protein sequences for Porphyra yezoensis were screened for potential allergens using AlgPred 2.0 and AllergenOnline, followed by the FAO allergenicity test using Allermatch. Data from this were used to predict the B-cell epitopes using the IEDP prediction tool and T-cell epitopes using MHC2Pred and were modeled using SWISS-MODEL and PyMOL to highlight specific epitopes. These models were assessed for quality using Global Quality Model Estimate (GQME) scores, ERRAT scores, and VERIFY 3D. Results showed fourteen (14) potential red seaweed allergens, four (4) of which were found in Kappaphycus alvarezii and ten (10) in Porphyra yezoensis. Several proteins of red seaweeds shared structural similarities with species normally associated with food allergies, such as common hazel, Atlantic salmon, and shark catfish, as well as other types of allergens such as those in house-dust mites, that could potentially induce cross-reactivity. Additionally anticipated were specific B-cell and T-cell epitopes and their specific peptide sequences that were incorporated in the 3D models, which were created for further comparison with other molecular structures of recognized allergens. Almost all of the 3D models had a GQME score of above 0.7 and had a high ERRAT score for overall quality but some failed to pass the VERIFY 3D test. This study could serve as a preliminary yet robust approach to identifying allergenic proteins in red seaweed and narrowing down potential existing cross-reactive allergens from various species that could aid in future in vitro and in vivo allergenicity studies.

Shotgun proteomics characterization of potential allergens in dried and powdered krill and fresh and powdered whiteleg shrimp

A shotgun proteomic and in-silico analysis was performed to identify and characterize the proteomes and potential allergens in both processed and non-processed forms of krill (Acetes spp.) and whiteleg shrimp (Litopenaeusvannamei). The samples were analyzed using LC-MS/MS, and the identified proteins were analyzed for protein-protein interactions and Gene Ontology. Potential allergens were analyzed by Allermatch and predicted IgE epitope by AlgPred. The results revealed that processed krill and whiteleg shrimp had a higher number of identified proteins and potential allergens compared to their non-processed forms. The major functions of identified proteins were related to organelle organization, intracellular and cytoskeletal protein binding. Eleven potential allergens were identified in all samples, indicating common allergens in both species. In-silico analysis of potential allergens of krill and whiteleg shrimp showed IgE epitopes on alpha-actinin sarcomeric-like isoform X1, tropomyosin fast isoform and tropomyosin isoform X19. The peptide IQLLEEDLER in tropomyosin fast isoform was identified as a potential allergen marker in fresh, dried and powdered forms of krill and whiteleg shrimp. These findings provide insight into the food safety, clinical and diagnostic aspects of krill and shrimp, particularly regarding its potential allergenicity and its allergen biomarkers.

INMUNOCAT study: The impact of molecular diagnosis on immunotherapy prescription in pollen polysensitized patients from Catalonia

Recognition of specific allergens triggering immune response is key for the appropriate prescription of allergen-specific immunotherapy (SIT). This study aimed at evaluating the impact of using the commercially available microarray ImmunoCAPTM ISAC 112 (Thermo Fisher Scientific) on the etiological diagnosis and SIT prescription compared to the conventional diagnostic methods in patients with allergic rhinitis/rhinoconjunctivitis and/or asthma. 300 patients with respiratory allergic disease, sensitized to three or more pollen aeroallergens from different species, as assessed by a skin prick test (SPT) and specific IgE assays (sIgE), were included in this multicentric, prospective observational study. SPT and a blood test were performed to all patients. Total serum IgE and sIgE (ImmunoCAPTM) for allergens found positive in the SPT and sIgE allergen components (ImmunoCAPTM ISAC 112) were measured. According to SPT results, the most prevalent pollen sensitizers in our population were Olea europaea followed by grass, Platanus acerifolia and Parietaria judaica. The molecular diagnosis (MD) revealed Ole e 1 as the most prevalent pollen sensitizer, followed by Cup a 1, Phl p 1, Cyn d 1, Par j 2, Pla a 1, 2, and 3 and Phl p 5. Immunotherapy prescription changed, due to MD testing, in 51% of the cases, with an increase of prescription of SIT from 39% to 65%. The identification of the allergen eliciting the respiratory disease is essential for a correct immunotherapy prescription. The advances in allergen characterization using methods, such as the commercial microarray ImmunoCAPTM ISAC 112, can help clinicians to improve SIT prescription.

House dust mite allergy: The importance of house dust mite allergens for diagnosis and immunotherapy

House dust mite (HDM) allergy belongs to the most important allergies and affects approximately 65–130 million people worldwide. Additionally, untreated HDM allergy may lead to the development of severe disease manifestations such as atopic dermatitis or asthma. Diagnosis and immunotherapy of HDM allergic patients are well established but are often hampered by the use of mite extracts that are of bad quality and lack important allergens. The use of individual allergens seems to be a promising alternative to natural allergen extracts, since they represent well-defined components that can easily be produced and quantified. However, a thorough characterization of the individual allergens is required to determine their clinical relevance and to identify those allergens that are required for correct diagnosis of HDM allergy and for successful immunotherapy. This review gives an update on the individual HDM allergens and their benefits for diagnosis and immunotherapy of HDM allergic patients.

Wood dust and asthma.

Review recent developments on asthma associated with wood dust, given the increasing scale of wood handling and processing activities globally. Work in wood industries is associated with a significantly increased risk of respiratory symptoms, rhinitis and asthma. This can be attributed to traditional processing techniques and newer technologies producing complex bioaerosol exposures, which may include chemicals. Meta-analysis studies indicate strong evidence for wood dusts as occupational sensitizers for asthma, but the underlying mechanisms remain poorly understood. The global prevalence of asthma in wood workers ranges between 6-18% and for rhinitis 16-33%. Exposure estimates show wide variation. Risk factors include atopy and exposure to certain wood species, elevated current and cumulative particulate exposures. Future studies should focus on better characterization of wood dust allergens and other bioaerosol components, specific immunoglobulin E responses to different wood species, pathophysiological mechanisms underlying asthma, and modelling dose-response relationships using refined exposure metrics for dust particulate and other bioaerosol components. There is a need for improved health-based international exposure standards and effective workplace control measures to reduce exposures to wood dust particulate (hard and soft woods), endotoxin and β-glucan, to reduce the risks of asthma in wood workers.

Molecular and immunological characterization of cysteine protease from Phaseolus vulgaris and evolutionary cross-reactivity.

A commonly consumed legume in India, the kidney bean (Phaseolus vulgaris) is associated with allergy. We report molecular and immunological characterization of cysteine protease allergen and its cross-reactivity. In silico allergenicity assessment and phylogenetic analysis of kidney bean cysteine protease showed significant sequence homology (upto 67%) with allergens from kiwi, papaya, soybean, ragweed pollen and mites. Physicochemical properties and motif-analysis depicted cysteine protease as probable allergen. Multiple sequence alignment and phylogenetic analysis indicated structural conservation between kidney bean and homologous cysteine protease sequences. The gene was cloned, expressed and affinity purified. Cysteine protease was resolved at 42 kDa and exhibited high IgE binding (up to 89%) with hypersensitive sera. Cysteine protease showed functional property on cross-linking IgE receptors and upregulated expression of CD203c on activated basophils. In inhibition studies, 8.4ng of cysteine protease was required for 50% self-inhibition, whereas significant inhibition was also observed with kidney bean (52 ng), black gram (155 ng), chick pea (437 ng), mesquite pollen (36 ng), house dust mite (64.85 ng), Alternaria alternata (78.8ng) and Curvularia lunata (73.6ng) extracts. ConSurf analysis indicated conserved active site and catalytic residues in mature domain among proteases from legumes, fruits, pollens, mites and fungus. In summary, P. vulgaris cysteine protease was molecularly characterized having functional activity. This study demonstrated, cross-reactivity between food and aeroallergens based on evolutionary conservancy that showed its clinical importance as cross-reactive allergen. PRACTICAL APPLICATIONS: Adaptation of sustainable lifestyle has led to a surge in consumption of plant-based foods especially legumes. Their high nutritional content lowers the risk of developing cardiovascular diseases, diabetes, obesity, and stroke. Kidney beans, a commonly consumed legume in Indian subcontinent, have a potential to be used as nutraceutical and functional food. Despite its alimentary nature, it elicits allergic reactions. Being a major sensitizer, trivial information regarding its allergic components has led to an urgent need for exploring its allergen repertoire. Our study reported biochemical and immunological characterization of its major cysteine protease allergen. Cysteine proteases are major cross-reactive allergens from insects, fruits and fungal sources. Identification and molecular characterization of such immunodominant allergens by RDT offers the prospect of using recombinant proteins for accurate diagnosis and therapeutic purposes. This study suggests that a potential major cross-reactive allergen may aid in developing allergy management interventions for a wide range of allergenic sources.

Storage Mite Precision Allergy Molecular Diagnosis in the Moderate-to-Severe T2-High Asthma Phenotype.

Storage mites (SM) may induce allergic respiratory symptoms in sensitized individuals, in both rural and urban settings. The relationship among specific IgE reactions to determined groups of SM allergens in the coincident asthma pheno-endotypes has not yet been investigated. We aimed to study a Precision Allergy Molecular Diagnosis (PAMD@) model to depict the SM molecular profile in individuals presenting with Type-2 inflammation, in two different (moderate and severe) asthma phenotypes. A customized PAMD@ panel, including SM allergens and their concurrent protein allergenic characterization was investigated. Mite group 2 allergens were most frequently recognized, including Lep d 2 (83.45%), followed by Gly d 2 (69.17%) and Tyr p 2 (47,37%), in 133/164 asthmatic subjects. Blo t 5 and Blo t 21 exhibited significant higher titres in both asthma groups. Although relevant mite group 2 allergens cross-reactivity is suggested, individualized sensitization patterns were relevantly identified. The present PAMD@ panel confirmed the dominance of mite group 2 allergens in moderate-to-severe T2 asthmatics. A broadly heterogeneous molecular repertoire of SM allergens was found in all subjects, regardless of their asthma severity. Blomia tropicalis deserves special attention in certain territories, as diagnostic and/or therapeutic approaches merely based on Pyroglyphidae mites may be insufficient.

A Review of Naturally Occuring Food Allergens and Their Impact on Health

Allergic reactions to foods influence a remarkable amount of population and are related with significant wellbeing results. It is one of the most significant issues that have expanding consideration. Current comprehension of the turn of events and utilization of allergenic capability of certain types of food proteins is restricted. In spite of the fact that there is a selection of in vivo models for assessing hypersensitivity, none of the current models has been approved, is prescient, or generally acknowledged with respect to their allergen explicit inhibitors. Hence, there is a proceeded with enthusiasm on the knowledge recovery based on food allergy so as to give more enlightening way to the current research field. In this paper, the current status of purification, characterization, and types of food allergens and their impacts is thoroughly reviewed. The present available methods for the allergen assessment (in view of animal, cell and clinical methodologies) are emphasized.

Allergenic Characterization of Best-Selling Hair Products Marketed to Black Women.

Overview | The leading peer-reviewed scientific journal in contact dermatitis, and the only journal in its field that publishes leading research in atopic, contact, occupational, environmental, and drug dermatitis.

House Dust Mite Precision Allergy Molecular Diagnosis (PAMD@) in the Th2-prone Atopic Dermatitis Endotype.

Atopic dermatitis (AD) endotyping might be important for developing personalized diagnostic and therapeutic strategies to the different phenotypes. The current study investigated the IgE molecular profile to Dermatophagoides pteronyssinus (D. pteronyssinus) in a subset of patients afflicted with varying severity stages of atopic dermatitis in a subtropical region subjected to a high perennial house dust mite (HDM) exposure. We selected patients showing a clinically relevant sensitization to HDM with mild-to-moderate and severe AD according to their basal Severity Scoring Atopic Dermatitis (SCORAD) index. Skin prick test (SPT) with standardized mite extracts, as well as a Precision Allergy Molecular Diagnosis (PAMD@) panel including nine different D. pteronyssinus allergens and the related protein allergenic characterization, were assessed in all serum samples. A total of 80 European American AD patients with the marked T2 endotype confirmed their eligibility for the study. Major allergens (Der p 23, Der p 2, and Der p 1) were present in more than 86% of all subjects, with mid-tier allergens (Der p 5, Der p 7, and Der p 21) reaching up to 65%. A serodominant role for Der p 11 could not be quantitatively confirmed in the present cohort. The proposed component resolved diagnosis (CRD) panel appeared to be sufficient to obtain a precise D. pteronyssinus molecular diagnosis in AD patients subjected to a climate-dependent high-mite allergen exposure. The raised seroprevalence of IgE response to Der p 23 confirmed this constituent as a major D. pteronyssinus allergen in severe stages of atopic dermatitis. A clinically driven molecular approach appears to be essential to frame a more precise diagnosis and therapy of this heterogeneous allergic condition.

Editorial introductions.

Current Opinion in Allergy and Clinical Immunology was launched in 2001. It is one of a successful series of review journals whose unique format is designed to provide a systematic and critical assessment of the literature as presented in the many primary journals. The fields of allergy and clinical immunology are divided into 14 sections that are reviewed once a year. Each section is assigned a Section Editor, a leading authority in the area, who identifies the most important topics at that time. Here we are pleased to introduce the Journal's Section Editors for this issue. SECTION EDITORS M. Teresa (Maite) de la MorenaM. Teresa (Maite) de la MorenaM. Teresa (Maite) de la Morena is Professor of Pediatrics in the Department of Pediatrics, Division of Immunology at University of Washington and Seattle Children's Hospital, USA. She graduated from medical school at Universidad Complutense de Madrid, Spain. This was followed by residency and fellowships training in pediatrics, allergy and immunology, and laboratory immunology at the University of South Florida, St. Petersburg, Florida, USA. Dr de La Morena has a clinical interest in allergy and immunology, clinical immunology with particular focus on the primary immunodeficiency diseases. Her research interests include long term clinical outcomes of patients with X-linked Hyper IgM syndrome, DiGeorge syndrome and microRNAs in primary immunodeficiency disorders, including their role in thymic development. Stephen JollesStephen JollesDr Jolles is a Consultant Clinical Immunologist and Honorary Professor at the University Hospital of Wales, UK. He is Clinical Lead in the Immunodeficiency Centre of Wales and Co-Director of the Hub Immunology Diagnostic Laboratory of Wales. Dr Jolles trained in immunology and allergy, and obtained his PhD on Langerhans cell migration at the National Institute for Medical Research, London. He has a particular interest in the clinical use of immunoglobulin and its mechanism of action and research interests include development of new therapeutic products, new screening strategies to improve the laboratory diagnosis of primary and secondary immunodeficiency and allergy, and better ways to monitor primary immunodeficiency disease and the impact of treatments. Giovanni PassalacquaGiovanni PassalacquaProf. Giovanni Passalacqua was born in Genoa in 1964, graduated in 1990 from the Genoa University Medical School, Italy, and was appointed by the specialty degrees in Allergy and Clinical Immunology and Respiratory Medicine in 1994 and 2002. He is the Co-Chairman of the Immunotherapy Scientific Committee of the WAO and member of the ARIA editorial board. Prof. Passalacqua serves as a section editor for Current Opinion in Allergy and Clinical Immunology, and also as a reviewer for Allergy, JACI, Clin Exp Allergy and ERJ. Prof. Passalacqua is the co-author of several international position papers, including ARIA guidelines, WAO Pos pap on SLIT, EAACI Pos Pap on local immunotherapy. His main fields of research are sublingual immunotherapy, mechanisms of allergic inflammation, antihistamines, corticosteroids, respiratory allergy, hymenoptera venom allergy. Under the chairmanship of Professor Canonica, he developed and investigated sublingual immunotherapy in the last 20 years. He has had 450 full articles published in peer reviewed journals indexed in MedLine plus 15 chapters in international handbooks. He currently heads the Allergy and Respiratory Diseases Department, and chairs the Specialty School in Respiratory Diseases, University of Genoa, Italy. Robert BushRobert BushDr Robert Bush received his MD degree from West Virginia University, USA. After completing his residency in internal medicine and fellowship in allergy and immunology at the University of Wisconsin – Madison, USA, he joined the faculty. He is currently Professor (Emeritus) of Medicine and was the former Chief of Allergy at the William S Middleton VA Hospital, Madison, Wisconsin, USA. His research interests include sublingual immunotherapy, allergen characterization, fungal allergy, food allergy, and occupational asthma.